Steroid 6-acetic acids



Patented Jan. 29, 1952 UNITED STATES PATENT OFFICE s'rnnoro fi-ACETICACIDS Emil Kaiser and Jerry Svarz, Chicago, Ill., as-

signors to Armour and Company, Chicago, 111., a corporation of IllinoisNo Drawing. Application May 8, 1948, Serial No. 26,002

1s Claims. (c1. zen-291.1)

method organic sulfonic esters of cholesterol are reacted with 000R HM-C ooon where M is a metal, and the reaction product is shown to be anisomeric mixture of cholesterol malonic esters. By saponification of theester mixture, a mixture of acids is obtained which can be separatedinto a petroleum ether insoluble fraction (Fraction A) and a petroleumether soluble fraction (Fraction B).

In our co-pending application, Serial No. 729,882, now Patent No.2,520,726, we have described Fraction B as a mixture which can itself beseparated into different fractions. A first fraction can be crystallizedfrom a' solution of Fraction B in methanol, and from the mother liquorof the methanol crystallization, a second fraction may be isolated incrystalline form. This second fraction is i-cholesterylmalonic acid.Methods of preparing this acid and its ester and amide derivatives aredescribed in said application, Serial No. 729,882.

We have now found that cyclopentanoperhydrophenanthrene compounds ingeneral are adaptable for use in the processes described above and thatit is possible to produce, for example, i-stigmasteryl-malonic acid,i-sitosterylmalonic acid, i-sitosterylmalonic acid esters,i-stigmasterylmalonamide, and-the like. Such compounds may be describedas cyclopentanoperhydrophenanthrene compounds of the i-form havingattached at their 6-position a malonic acid group, or a malonic acidester group, or a malonamide group, etc. i

We have found further that the above mal-onic acid compounds may betreated to produce, as new compositions of matter,cyclopentanoperhydrophenanthrene compounds of the i-form, havingattached at their fi-position an acetic acid group. Such treatmentcomprises (1) decarboxylation at elevated temperatures in high vacuum;or (2) decarboxylation by conversion to the amide through'use of"thionyl chloride and ammonia, followed by saponification of the amidewith an alkaline glycol.

The following equations are illustrative of the two reactions, where thecyclopentanoperhydro- 5 phenanthrene compound is a cholesterolderivative:

CH; CH; cm

j 47H: A decarboxylatlon at elevated v temperature in high vacuum C O OHi-Choles terylmalonic acid CH3 on, on, H--(CH2)9(H Hz O Oi H1 0 0 oni-Cholesterylacetlc acid (M. r. 111-112" li?+ CH3 CH3 CH, BP-(CHQF- BH CH:

thionylchlorlde ammonia O 0 0E H C O OH l-Cholesterylmalonlc acidsaponification with alkaline glycol L/ H10 ZNBs CO:

Preparation ofi-chole sterylc c etic acid from icholesteryl mclonic acidIn a 500 cc. flask immersed in an oil bath, heat 10 g. ofi-cholesterylmalonic acid in a vacuum maintained by a mercury vapor.pump. Raise the temperature slowly to 175 and keep it there for 10minutes. Then slowly] raise the temperature again to 200 and continueheating for 10 more minutes. After that, cool the flask to roomtemperature and crystaliize the reaction product from acetone. Becrystall'ze from acetone to obtain the i-cholesterylacet c acid.

EXAMIQLEII Preparation of i-stigmasterylmc lonamide In 10 cc. ofthionylchloride, suspend 5 g. of istigmasterylmalonic acid andlgeepthemixture at room temperature for two hours. Bring the material whichis still undissolved into solution by heating on the steam bath forlfllminutes.

Distill off the excess thionylchloride under re duced pressure and takeup the crude acid chloride in 75 ccl'of absolute ether. Filter the ethersolution and pass dry ammonia gas into thefiltrate for 70 minutes.Remove the ether and wash the residue with water. After drying in vacuo,crystallize the i-stigmasterylmalonamide. from ethyl acetate.

EXAMPLE III Saponification of i-stigmasterylmalonamide Dissolve 20 g. ofpotassium hydroxide by warming in 100 cc. of propylene glycol. Add 7 g.of istigmasterylmalonamide and reflux the mixture ior 10 hours.Saponification 9f the amide will be indicated by the evolution ofammonia during the heating; Dilute the clear propylene glycol solutionwith water, filter, and acidity with hy drochloric acid. Extract theresulting precipitate with other Evaporate the ether solution, dissolvethe residue in methanol, and treat the solution with charcoal. Afterfiltration and chilling, needles will he obtained which may be twicerecrystallized from methanol. The resulting produc t isistigrnasterylacetic acid.

We have found further that the cyolopentanoperhydrophenanthrenecompounds of the iform, having a malonamide or an acetic acid groupattached at their 6-position, may be treated with a mineral acid wherebyto effect re rrangement and produce, as newcompositions matter,unsaturated cyclopentanoperhydrophenanthrone compounds having an aceticacid group eq org 'cai ieles se rfi-e ci s; a id; (M..- B, 13. 1 answe Te m th r liqu r mainin a r salv tio o e hole ene-fi-arotic ac d, Qetaiew r a e ton. ringontain ng p oduct w i h i also new and which has thestructural formula;

Si ar y. whe -i h stc ylm o mies., for

example, is dissolved in glacial acetic acid ree ad th. qoncea reiccl eli t c. a id a. ro.

acetic acid, the structure of which is pictured above. The mother liquorremaining after separation of the cholestene-fi-acetic acid-contains thesame neutral, lactone ring-containing product, the structure of which isalso pictured above.

Any mineral acid may be used in bringing about rearrangement. Examplesof such acids are sulfuric acid, phosphoric acid, nitric acid, and thelike.

By application of the foregoing procedures, the new rearrangementproducts comprising unsaturated cyclopentanoperhydrophenanthrenecompounds having an acetic acidgroup attached at their G-position andcyclopentanoperhydrophenanthrene compounds in which the carbon atomsoccupying the and 6-positions form part of a lactone ring may beprepared by starting with a cyclopentanoperhydrophenanthrene compound ofthe i-form having attached at its 6-position a malonamide group andsubjecting it to treatment comprising (1) saponiiication with alkalineglycol, followed by acid rearrangement; or (2) acid rearrangement,followed by saponification with alkaline glycol.

These new rearrangement products are useful as intermediates in thepreparation of vitamins and hormones of the progesterone type and malehormone type. The following examples serve to illustrate the manner inwhich they may be prepared.

EXAMPLE IV Rearrangement of i-cholesterylacetic acid Dissolve 4 g. ofi-cholesterylacetic acid in 120 cc. of glacial acetic acid. Add drops ofconcentrated sulfuric acid and heat the solution on the steam bath fortwo hours. Cool the reaction mixture and pour into one liter of 10%aqueous sodium chloride solution. Filter oil the precipitate, dry, anddissolve in 20 cc. of glacial acetic acid. After filtration, keep thesolution at room temperature for several days. Filter to remove thecrystals which deposit. Concentrate the mother liquor to half its volumeunder reduced pressure to obtain a second crop of crystals. Thesecrystals are cholestene-G-acetic acid.

After the cholestene-G-acetic acid is crystallized out from theconcentrated glacial acetic acid solution, evaporate the mother liquorto dryness under reduced pressure. Dissolve the oily residue in etherand repeatedly extract with a 5% potassium hydroxide solution. Thecombined alkaline extracts will yield. additional amounts ofcholestene-G-acetic acid.

Wash the ether layer with water, dry over sodium sulfate, and evaporatethe ether. The neutral. lactone ring-containing rearrangement productWill crystallize in beautiful needles.

EXAMPLE v Rearrangement of i-sitosterylmalonamide i-Sitosterylmalonamidemay be prepared in a process similar to that described in Example IIwhich relates to the preparation of i-stigmasterylmalonamide. In 200 c.c. of glacial acetic acid, dissolve 2.25 g. of thei-sitosterylmalonamide thus prepared and add 1 c. c. of concentratedsulfuric acid. Keep the mixture on the steam bath for one hour, then add500 c. c. of water. Filter off the precipitate, wash with water, anddry. Dissolve the dry material in c. c. of methanol and keep thesolution at --4 for 12 a hours. Filter to separate the swollen lumpswhich deposit. After drying. a slightly colored powder'will be obtainedwhich is soluble in ether and benzene and which may be designated asFraction I.

Dilute the mother liquor filtered oil from Fraction I with water. Awhite precipitate will form, which may be designatedFraction II.

EXAMPLE VI Hydrolysis 0 Fraction I Dissolve 1 g. of potassium hydroxidein 10 c. c. of propylene glycol and add 0.50 g. of Fraction I. Refluxthe solution for two hours, cool and dilute ,with c. 0. water. Filteroff the precipitate,

wash with water, and dry. crystallize the dry material from glacialacetic acid. Recrystallize from methanol to obtain a pure form of therearrangement product having the following structure:

(tin -coon EXAMPLE VII Hydrolysis of Fraction II Reflux 1.5 g. ofFraction II in 25 c. c. of propylene glycol containing 2.5 g. ofpotassium hydroxide. After 2 hours refluxing, cool the solu-- tion,dilute with 400 c. c. of water (clear solution) and acidify. Filter offthe precipitate, wash with water, and dry. Dissolve the dry material in20 c. c. of methanol. After dilution with 100 c. c. of water, extractthe mixture with petroleum ether. Evaporate the petroleum ether.Rec'rystallize the residue twice from methanol. The resulting needlescomprise the lactone ringcontaining rearrangement product having thefollowing structure:

The foregoing examples indicate certain specific ways in which ourimprovements may be practiced-but they are given for the purpose ofexplanation only, and it is understood. that many other specificprocedures may be used, all within the spirit of the invention.

We claim: '1. A cyclopentanoperhydrophenanthrene compound havingattached at its S-position an acetic acid group.

2. Cholestene-6-acetic acid.

A compound mag the structure CH; @135 GB:

. fi-position a malonic acid group, separating from the reaction mixturethe resulting malonamide, saponifying said malonanlide compound with analkaline glycol, acidifying the resulting solution, and separating fromsaid solution a precipitate comprising acyclopentanoperhydrophenanthrene compound of the i-form having'attachedat its (i-position an acetic acid group.

9. A process ior theproduction of a cyclopentanoperhydrophenanthreriecompound comp sin apo ifyiue. w th alkali e ycol acyclopcntanoperhydrophenanthrene compound of t -ferm ine t ched at it -Pion a malonan ide group, acidifying the resulting soiution, andseparating from said solution a precip t omp sing a cyc pentao peydronan e c mp nd f th rm h i attached at its S-p sition an acetic acidgroup.

10. A process for the production of i-cho'lesterylacetic acid comprisingreacting i-ch'olesteryb r lou acid, i th omr o r de and a monia,

p rat ng i h sterrlma o amide rom the resulting reaction 1 ture,saponifying said i-cholesterylmalonamide with, alkaline glycol,

acidifying the resulting solution, and'separating i-cholesterylaceticacid in the form of a precipitate from said solution.

11. A p f-Qcess for producing a cyclopentanoperhydrophenanthrenecompound comprising heating under reduced pressure acyclopentanoperhydrophenanthrene compound of the i-form having attachedat its .Q-position a malonic acid group, cooling the reaction mixture,and separating the precipitate comprising a cyclopentanoper"hydrophenanthrene compound of the i-iorm having an acetic acid groupattached at its 6-position.

12. A process for the production of i-cholesterylacetic acid comprisingheating i-cholesterylmalonic acid under reduced pressure, cooling, andseparating .i-cholesterylacetic acid in the f m of a p e p ta e mthereac i n mixturea p oce s for r duc ng mam-rated cyclo entanop rhd-ropheuau reu oomuund om ng treati a eture edoy lopent-aonerhydrophenanthrene compound ,of 1 1e i-iorm, having attached at its6-position an, acetic, acid gro p, With a mineral acidwhereby to fi c area rangement eml reti s the rea t on mirture with an aqueous sod umchloride solution to roduce a precip ta e omp is ng an n aturatedcyclopentancperhydrO henanthrene compound having attached at itsdeposition an acetic a d group. 7 r

1-11, A rocess for producing an unsaturated cyclopentanop h r ph n nthrne c mp un compr sms P ng n. a rea tio essel a sa ratedcyclopentanoperhydrophenanthrene comratedcyclopentanoperhydrophenanthrene com-- pound comprising reacting withthionylchloride and ammonia a saturated cyclopentano erhydrophenanthrene-.compound or" the, ieforsn havi g attached at its G-position a maloniacid group, separating from the reaction mixture, the result, mgmalonamide, treating said ,maionamide (59m?- pound With sulfuric acidwhereby t produce a rearrangement product, cooling a methanol solu-,tion of said rearrangement product until a prepi ate is for ed, s po iyi sa d prec pita e with an alkaline glycol solution, and separatingfrom the reaction mixture an unsaturated cyclepentanoperhydrophenanthrene compound having attached at its d-positionan acetic acid group.

16. A process for the production of an unsaturatedcyclopentanoperhydrophenanthrene iconiound comprising sapcnifyir g withan aiigaline the resulting solution, separating from said solution theresulting acetic acid compound, treating said compound with sulfuricacid whereby to eifect a rearran ement, {and treating. the reaction migture with an aqueous sodium chloride soluti n. to prod a prec p tatempr si a unsaturated .cyclopentan p rhrdroshen n hr e compou d h v ngatt ch d at its fi-position an acetic acid eroun- 171- A pr cess o thproducti n o ohole t l vfi-e aci comp is n t ating iro l s er l m lonamde wit su f r c acid ereb t pro: duce a rearrangement. product, coolinga nethanol solution of said rearrangement product t l a pre tate is io eseo ni y ne a p cipitate with an alkaline glycol solution, andseparating cholestfllQ-fiemetic acid from the react o mixture. 7

v1.3- A prooess. for h p oduction of cholestene- G-acetic acidcomprising saponiiying i-cholesterylmalonamide with an alkaline glycol,acidifying the resulting solution, separating i-choI- esterylacetic acidin. the :form of a precipitate from said solution, treati ofe Said,i-choleeterylacetic acid with sulfuric acid whereby to effect arearrangement, treating the reaction mixture w th aqueous sodiumchloride solution to, produce, ChOlGStEDG-B-1&C Qti0 acid the form aprecipi ate.

Ell EL KAISER.

JERRY SVARZ.

0 refer nce Lei...

1. A CYCLOPENTANOPERHYDROPHENANTHRENE COMPOUND HAVING ATTACHED AT ITS6-POSITION AN ACETIC ACID GROUP.